U.S. patent number 6,166,053 [Application Number 09/490,269] was granted by the patent office on 2000-12-26 for substituted condensation products of n-benzyl-3-idenylacetamides with heterocyclic aldehydes for neoplasia.
This patent grant is currently assigned to Cell Pathways, Inc.. Invention is credited to Klaus Brendel, Paul Gross, Rifat Pamukcu, Gary A. Piazza, Gerhard J. Sperl.
United States Patent |
6,166,053 |
Sperl , et al. |
December 26, 2000 |
Substituted condensation products of N-benzyl-3-idenylacetamides
with heterocyclic aldehydes for neoplasia
Abstract
Substituted condensation products of
N-benzyl-3-indenylacetamides with heterocyclic aldehydes are useful
for inducing or promoting apotosis and for arresting uncontrolled
neoplastic cell proliferation, and are specifically useful in the
arresting and treatment of neoplasias, including precancerous and
cancerous lesions.
Inventors: |
Sperl; Gerhard J. (North Wales,
PA), Gross; Paul (Stockton, CA), Brendel; Klaus
(Tuscon, AZ), Piazza; Gary A. (Doylestown, PA), Pamukcu;
Rifat (Spring House, PA) |
Assignee: |
Cell Pathways, Inc. (Horsham,
PA)
|
Family
ID: |
26901185 |
Appl.
No.: |
09/490,269 |
Filed: |
January 24, 2000 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
Issue Date |
|
|
206245 |
Dec 7, 1998 |
6066634 |
|
|
|
989353 |
Dec 12, 1997 |
5948779 |
|
|
|
Current U.S.
Class: |
514/357;
546/333 |
Current CPC
Class: |
C07D
241/12 (20130101); A61P 35/00 (20180101); C07D
239/26 (20130101); C07D 215/14 (20130101); C07D
213/56 (20130101); C07D 237/08 (20130101) |
Current International
Class: |
C07D
215/00 (20060101); C07D 237/00 (20060101); C07D
237/08 (20060101); C07D 215/14 (20060101); C07D
241/00 (20060101); C07D 213/00 (20060101); C07D
241/12 (20060101); C07D 239/26 (20060101); C07D
213/56 (20060101); C07D 239/00 (20060101); A61K
031/4409 (); C07D 213/56 () |
Field of
Search: |
;514/357 ;546/333 |
References Cited
[Referenced By]
U.S. Patent Documents
|
|
|
5948779 |
September 1999 |
Sperl et al. |
|
Primary Examiner: Raymond; Richard L.
Assistant Examiner: Rao; Deepak R.
Attorney, Agent or Firm: Stevenson; Robert W.
Parent Case Text
This application is a continuation of U.S. patent application Ser.
No. 09/206,245, filed Dec. 7, 1998 now U.S. Pat. No. 6,066,634
which is a continuation-in-part of U.S. patent application Ser. No.
08/989,353 filed Dec. 12, 1997 now U.S. Pat. No. 5,948,779.
Claims
We claim:
1. A method of treating a patient having neoplasia comprising
administering a pharmacologically effective amount of
(Z)-5-fluoro-2-methyl-(4-pyridylidene)-3-(N-benzyl)indenylacetamide
hydrochloride to the patient with a neoplasia sensitive to such a
compound.
2. A method for regulating apoptosis in human cells comprising
exposing the cells to an effective amount of
(Z)-5-fluoro-2-methyl-(4-pyridylidene)-3-(N-benzyl)indenylacetamide
hydrochloride.
Description
TECHNICAL FIELD
This invention relates to compounds and methods for inducing or
promoting apotosis and for arresting uncontrolled neoplastic cell
proliferation, methods that are specifically useful in the
arresting and treatment of neoplasias, including precancerous and
cancerous lesions.
BACKGROUND OF THE INVENTION
Pharmaceuticals that are effective against early stage neoplasias
comprise an emerging and expanding area of research and potential
commercial development. Such pharmaceuticals can delay or arrest
development of precancerous lesions into cancers. Each year in the
United States alone, untold numbers of people develop precancerous
lesions, which exhibit a strong statistically significant tendency
to develop into malignant tumors, or cancer. Such lesions include
lesions of the breast (that can develop into breast cancer),
lesions of the skin (that can develop into malignant melanoma or
basal cell carcinoma), colonic adenomatous polyps (that can develop
into colon cancer), cervical dysplasia (cervical cancer) and other
such neoplasms.
Compounds that prevent or induce the remission of existing
precancerous or cancerous lesions, or carcinomas, delay the onset
of cancer and would greatly reduce illness and death from at least
certain forms of that disease.
Such compounds and methods are particularly beneficial to
sub-populations of patients who repeatedly develop precancerous
lesions, and therefore have a statistically higher probability of
getting cancer. Many cancer types (e.g., breast, colon, prostate
etc.) have such patient sub-populations. One example of a
sub-population that will invariably develop cancer (if left
untreated) includes those patients who suffer from familial
polyposis of the colon. Familial polyposis patients typically
develop many (e.g., hundreds or thousands) of colonic polyps
beginning in their teenage years. Because each colonic polyp
(whether familial or non-familial) reportedly has approximately a
five percent lifetime risk of developing into a cancer, the
treatment of choice--until very recently--for familial polyposis
patients is surgical removal of the colon in the early
twenties.
Many other cancers have sub-populations that also have much higher
risks for getting cancer at an early age and for having the cancer
reoccur, than patients as a whole who get such a cancer. For
example, such sub-populations have been identified among breast
cancer patients and colon cancer patients. In the latter
sub-population, removal of the individual polyps as they form is
the current treatment of choice. Removal of polyps in non-familial
patients has been accomplished either with surgery or fiber-optic
endoscopic polypectomy--procedures that are uncomfortable, costly
(the cost of a single polypectomy ranges between $1,000 and $1,500
for endoscopic treatment and more for surgery), and involve a small
but significant risk of colon perforation.
The search for drugs useful for treating and preventing neoplasias
in their earliest stages is intensive because chemotherapy and
surgery on cancer itself is often not effective, and current
chemotherapy has severe side effects. Thus, the search for
compounds effective against precancerous lesions without the side
effects of conventional chemotherapy is particularly intensive.
Such compounds are also envisaged for recovered cancer patients who
retain a risk of cancer reoccurrence, and even for cancer patients
who would benefit from compounds that selectively induce apoptosis
in neoplastic, but substantially not in normal cells.
Standard cancer chemotherapeutic drugs are not considered
appropriate drugs for cancer chemoprevention because whatever
cancer preventative (as opposed to cancer-fighting) capabilities
those drugs may possess do not outweigh their severe side effects.
Most standard chemotherapeutics are now believed to kill cancer
cells by inducing apoptosis (also sometimes referred to as
"programmed cell death"). Apoptosis naturally occurs in virtually
all tissues of the body. Apoptosis plays a critical role in tissue
homeostasis, that is, it ensures that the number of new cells
produced are correspondingly offset by an equal number of cells
that die. Apoptosis is especially pronounced in self-renewing
tissues such as bone marrow, immune cells, gut, and skin. For
example, the cells in the intestinal lining divide so rapidly that
the body must eliminate cells after only three days to protect and
prevent the overgrowth of the intestinal lining.
Standard chemotherapeutics promote apoptosis not only in cancer
cells, but also in normal human tissues, and therefore have a
particularly severe effect on cells that normally divide rapidly in
the body (e.g. hair, gut and skin). The results of those effects on
normal cells include hair loss, weight loss, vomiting and bone
marrow immune suppression. This is one reason standard
chemotherapeutics are inappropriate for cancer prevention.
In the absence of a one-time cure (e.g., a gene therapy), another
reason is that cancer prevention therapy requires chronic
administration of a pharmaceutical to repress neoplasia formation,
which for standard chemotherapeutics is obviously contraindicated
because of the types of side effects discussed above.
Abnormalities in apoptosis can lead to the formation of
precancerous lesions and carcinomas. Also, recent research
indicates that defects in apoptosis play a major role in other
diseases in addition to cancer. Consequently, compounds that
modulate apoptosis could be used in the prevention or control of
cancer, as well as other diseases.
Several non-steroidal anti-inflammatory drugs ("NSAIDs"),
originally developed to treat arthritis, have shown effectiveness
in inhibiting and eliminating colonic polyps. Polyps virtually
disappear when the patients take the drug, particularly when the
NSAID sulindac is administered. However, the continued prophylactic
use of currently available NSAIDs, even in polyposis syndrome
patients, is still marked by severe side reactions that include
gastrointestinal irritations, perforations, ulcerations and kidney
toxicity believed to be produced by inhibition of prostaglandin
synthetase activity ("PGE-2"). Such inhibition is a requirement for
the NSAIDs anti-inflammatory action since elevated levels of PGE-2
are associated with inflammation. PGE-2 plays a protective function
in the gastrointestinal tract, which is the reason such gastric
side effects arise with chronic NSAID therapy, which is rarely
indicated for arthritis sufferers, acute therapy being the norm for
them. However, chronic administration of sulindac is important for
polyposis patients to eliminate and prevent future polyps which
causes gastric side effects in many such patients. Once NSAID
treatment is terminated due to such complications, the polyps
return, particularly in polyposis syndrome patients.
Compounds such as those disclosed in U.S. Pat. No. 5,643,959 have
exhibited advantages in the treatment of neoplastic lesions since
such compounds have been shown to induce apotosis in neoplastic
cells but not in normal cells in humans. Thus, the severe side
effects due to induction of apotosis in normal cells by
conventional chemotherapeutics are avoided by these novel
therapeutics (see, "Phase I Trial of Sulindac Sulfone in Patients
With Familial Polyposis (FAP) With Rectal Polyps: Optimal Dose and
Safety," Digestive Disease Week, Abstract No. 2457, May 10-16,
1997, American Gastroenterological Association et al.). In
addition, such compounds do not exhibit the gastric side effects
associated with NSAIDs since such compounds do not substantially
inhibit PGE-2. More potent compounds with such neoplasia
specificity but without substantial PGE-2 activity are
desirable.
SUMMARY OF THE INVENTION
This invention represents potent compounds, that induce apotosis in
neoplastic cells (but not substantially in normal cells), for
treating patients with neoplastic lesions without substantially
inhibiting PGE-2. This invention also involves methods for inducing
such specific apotosis in neoplastic cells by exposing such cells
to a pharmacologically effective amount of those compounds
described below to a patient in need of such treatment. Such
compositions are effective in modulating apoptosis and modulating
the growth of precancerous lesions and neoplasms, but are not
suffering from the side effects of conventional chemotherapeutics
and NSAIDs.
DETAILED DESCRIPTION OF THE INVENTION
As discussed above, the present invention includes compounds of
Formula I below (as well as their pharmaceutically acceptable
salts) for treating a patient with neoplastic, particularly
precancerous, lesions: ##STR1##
wherein R.sub.1 is independently selected in each instance from the
group consisting of hydrogen, halogen, lower alkyl, lower alkoxy,
amino, lower alkylamino, di-lower alkylamino, lower alkylmercapto,
lower alkyl sulfonyl, cyano, carboxamide, carboxylic acid,
mercapto, sulfonic acid, xanthate and hydroxy;
R.sub.2 is selected from the group consisting of hydrogen and lower
alkyl;
R.sub.3 is selected from the group consisting of hydrogen, halogen,
amino, hydroxy, lower alkyl amino, and di-loweralkylamino;
R.sub.4 is hydrogen, or R.sub.3 and R.sub.4 together are
oxygen;
R.sub.5 and R.sub.6 are independently selected from the group
consisting of hydrogen, lower alkyl, hydroxy-substituted lower
alkyl, amino lower alkyl, lower alkylamino-lower alkyl, lower alkyl
amino di-lower alkyl, lower alkyl nitrile, --CO.sub.2 H,
--C(O)NH.sub.2, and a C.sub.2 to C.sub.6 amino acid;
R.sub.7 is independently selected in each instance from the group
consisting of hydrogen, amino lower alkyl, lower alkoxy, lower
alkyl, hydroxy, amino, lower alkyl amino, di-lower alkyl amino,
halogen, --CO.sub.2 H, --SO.sub.3 H, --SO.sub.2 NH.sub.2, and
--SO.sub.2 (lower alkyl);
m and n are integers from 0 to 3 independently selected from one
another;
Y is selected from the group consisting of quinolinyl,
isoquinolinyl, pyridinyl, pyrimidinyl, pyrazinyl, imidazolyl,
indolyl, benzimidazolyl, triazinyl, tetrazolyl, thiophenyl,
furanyl, thiazolyl, pyrazolyl, or pyrrolyl, or subsituted variants
thereof wherein the substituents are one or two selected from the
group consisting of halogen, lower alkyl, lower alkoxy, amino,
lower alkylamino, di-lower alkylamino, hydroxy, --SO.sub.2 (lower
alkyl) and --SO.sub.2 NH.sub.2.
Preferred compounds of this invention for use with the methods
described herein include those of Formula I where:
R.sub.1 is selected from the group consisting of halogen, lower
alkoxy, amino, hydroxy, lower alkylamino and di-loweralkylamino,
preferably halogen, lower alkoxy, amino and hydroxy;
R.sub.2 is lower alkyl;
R.sub.3 is selected from the group consisting of hydrogen, halogen,
hydroxy, amino, lower alkylamino and di-loweralkylamino,
preferably, hydrogen, hydroxy and lower alkylamino;
R.sub.5 and R.sub.6 are independently selected from the group
consisting of hydrogen, hydroxy-substituted lower alkyl, amino
lower alkyl, lower alkylamino-lower alkyl, lower alkyl amino
di-lower alkyl, --CO.sub.2 H, --C(O)NH.sub.2 ; preferably hydrogen,
hydroxy-substituted lower alkyl, lower alkyl amino di-lower alkyl,
--CO.sub.2 H, and --C(O)NH.sub.2 ;
R.sub.7 is independently selected in each instance from the group
consisting of hydrogen, lower alkoxy, hydroxy, amino, lower alkyl
amino, di-lower alkyl amino, halogen, --CO.sub.2 H, --SO.sub.3 H,
--SO.sub.2 NH.sub.2, and --SO.sub.2 (lower alkyl); preferably
hydrogen, lower alkoxy, hydroxy, amino, amino lower alkyl, halogen,
--CO.sub.2 H, --SO.sub.3 H, --SO.sub.2 NH.sub.2, and --SO.sub.2
(lower alkyl);
Preferably, at least one of the R.sub.7 substituents is para- or
ortho-located; most preferably ortho-located;
Y is selected from the group consisting of quinolinyl,
isoquinolinyl, pyridinyl, pyrimidinyl and pyrazinyl or said
substituted variants thereof.
Preferably, the substituents on Y are one or two selected from the
group consisting of lower alkoxy, amino, lower alkylamino, di-lower
alkylamino, hydroxy, --SO.sub.2 (lower alkyl) and --SO.sub.2
NH.sub.2 ; most preferably lower alkoxy, di-lower alkylamino,
hydroxy, --SO.sub.2 (lower alkyl) and --SO.sub.2 NH.sub.2.
The present invention also is a method of treating a patient with
such lesions by administering to a patient a pharmacologically
effective amount of a pharmaceutical composition that includes a
compound of Formula I, wherein R.sub.1 through R.sub.7 and Y are as
defined above. Preferably, this composition is administered without
therapeutic amounts of an NSAID.
The present invention is also a method of treating individuals with
neoplastic lesions by administering a pharmacologically effective
amount of an enterically coated pharmaceutical composition that
includes compounds of this invention.
Also, the present invention is a method of inhibiting the growth of
neoplastic cells by exposing the cells to an effective amount of
compounds of Formula I, wherein R.sub.1 through R.sub.7 and Y are
defined as above.
In still another form, the invention is a method of inducing
apoptosis in human cells by exposing those cells to an effective
amount of compounds of Formula I, wherein R.sub.1 through R.sub.7
and Y are defined as above where such cells are sensitive to these
compounds.
Additionally, in yet another form, the invention is a method of
treating a patient having a disease which would benefit from
regulation of apoptosis by treating the patient with an effective
amount of compounds of Formula I, wherein R.sub.1 through R.sub.8
are defined as above. The regulation of apoptosis is believed to
play an important role in diseases associated with abnormalities of
cellular growth patterns such as benign prostatic hyperplasia,
neurodegenerative diseases such as Parkinson's disease, autoimmune
diseases including multiple sclerosis and rheumatoid arthritis,
infectious diseases such as AIDS, and other diseases, as well.
Compounds of this invention are also inhibitors of cGMP-specific
phosphodiesterase activity found in neoplastic cells. Such
phosphodiesterases include PDE5 as well as the novel PDE disclosed
in U.S. patent application Ser. No. 09/173,375 filed Oct. 15, 1998
to Pamukcu et al. For convenience, the PDE inhibitory activity of
such compounds can be tested as taught in co-pending U.S. patent
application Ser. No. 09/046,739 filed Mar. 24, 1998 to Pamukcu et
al., which is incorporated herein by reference. Thus, compounds of
this invention are useful inhibitors of PDE5 and may be useful in
medical indications where inhibition of that enzyme activity is
desired.
As used herein, the term "precancerous lesion" includes syndromes
represented by abnormal neoplastic, including dysplastic, changes
of tissue. Examples include dysplasic growths in colonic, breast,
bladder or lung tissues, or conditions such as dysplastic nevus
syndrome, a precursor to malignant melanoma of the skin. Examples
also include, in addition to dysplastic nevus syndromes, polyposis
syndromes, colonic polyps, precancerous lesions of the cervix
(i.e., cervical dysplasia), esophagus, prostatic dysplasia,
bronchial dysplasia, breast, bladder and/or skin and related
conditions (e.g., actinic keratosis), whether the lesions are
clinically identifiable or not.
As used herein, the term "carcinomas" refers to lesions that are
cancerous. Examples include malignant melanomas, breast cancer,
prostate cancer and colon cancer.
As used herein, the term "neoplasm" refers to both precancerous and
cancerous lesions and hyperplasia.
As used herein, the term "halo" or "halogen" refers to chloro,
bromo, fluoro and iodo groups, and the term "alkyl" refers to
straight, branched or cyclic alkyl groups and to substituted aryl
alkyl groups. The term "lower alkyl" refers to C.sub.1 to C.sub.8
alkyl groups.
The term "hydroxy-substituted lower alkyl" refers to lower alkyl
groups that are substituted with at least one hydroxy group,
preferably no more than three hydroxy groups.
The term "--SO.sub.2 (lower alkyl)" refers to a sulfonyl group that
is substituted with a lower alkyl group.
The term "lower alkoxy" refers to alkoxy groups having from 1 to 8
carbons, including straight, branched or cyclic arrangements.
The term "lower alkylmercapto" refers to a sulfide group that is
substituted with a lower alkyl group; and the term "lower alkyl
sulfonyl" refers to a sulfone group that is substituted with a
lower alkyl group.
The term "pharmaceutically acceptable salt" refers to non-toxic
acid addition salts and alkaline earth metal salts of the compounds
of Formula I. The salts can be prepared in situ during the final
isolation and purification of such compounds, or separately by
reacting the free base or acid functions with a suitable organic
acid or base, for example. Representative acid addition salts
include the hydrochloride, hydrobromide, sulfate, bisulfate,
acetate, valerate, oleate, palmatate, stearate, laurate, borate,
benzoate, lactate, phosphate, tosylate, mesylate, citrate, maleate,
fumarate, succinate, tartrate, glucoheptonate, lactobionate, lauryl
sulfate salts and the like. Representative alkali and alkaline
earth metal salts include the sodium, calcium, potassium and
magnesium salts.
It will be appreciated that certain compounds of Formula I can
possess an asymmetric carbon atom and are thus capable of existing
as enantiomers. Unless otherwise specified, this invention includes
such enantiomers, including any racemates. The separate enaniomers
may be synthesized from chiral starting materials, or the racemates
can be resolved by conventional procedures that are well known in
the art of chemistry such as chiral chromatography, fractional
cyrstallization of diastereomeric salts and the like.
Compounds of Formula I also can exist as geometrical isomers (Z and
E); the Z isomer is preferred.
Compounds of this invention may be formulated into pharmaceutical
compositions together with pharmaceutically acceptable carriers for
oral administration in solid or liquid form, or for rectal or
topical administration, although carriers for oral administration
are most preferred.
Pharmaceutically acceptable carriers for oral administration
include capsules, tablets, pills, powders, troches and granules. In
such solid dosage forms, the carrier can comprise at least one
inert diluent such as sucrose, lactose or starch. Such carriers can
also comprise, as is normal practice, additional substances other
than diluents, e.g., lubricating agents such as magnesium stearate.
In the case of capsules, tablets, troches and pills, the carriers
may also comprise buffering agents. Carriers such as tablets, pills
and granules can be prepared with enteric coatings on the surfaces
of the tablets, pills or granules. Alternatively, the enterically
coated compound can be pressed into a tablet, pill, or granule, and
the tablet, pill or granules for administration to the patient.
Preferred enteric coatings include those that dissolve or
disintegrate at colonic pH such as shellac or Eudraget S.
Pharmaceutically acceptable carriers include liquid dosage forms
for oral administration, e.g., pharmaceutically acceptable
emulsions, solutions, suspensions, syrups and elixirs containing
inert diluents commonly used in the art, such as water. Besides
such inert diluents, compositions can also include adjuvants such
as wetting agents, emulsifying and suspending agents, and
sweetening, flavoring and perfuming agents.
Pharmaceutically acceptable carriers for topical administration
include DMSO, alcohol or propylene glycol and the like that can be
employed with patches or other liquid-retaining material to hold
the medicament in place on the skin so that the medicament will not
dry out.
Pharmaceutically acceptable carriers for rectal administration are
preferably suppositories that may contain, in addition to the
compounds of this invention excipients such as cocoa butter or a
suppository wax, or gel.
The pharmaceutically acceptable carrier and compounds of this
invention are formulated into unit dosage forms for administration
to a patient. The dosage levels of active ingredient (i.e.,
compounds of this invention) in the unit dosage may be varied so as
to obtain an amount of active ingredient effective to achieve
lesion-eliminating activity in accordance with the desired method
of administration (i.e., oral or rectal). The selected dosage level
therefore depends upon the nature of the active compound
administered, the route of administration, the desired duration of
treatment, and other factors. If desired, the unit dosage may be
such that the daily requirement for active compound is in one dose,
or divided among multiple doses for administration, e.g., two to
four times per day.
The pharmaceutical compositions of this invention are preferably
packaged in a container (e.g., a box or bottle, or both) with
suitable printed material (e.g., a package insert) containing
indications, directions for use, etc.
There are several general schemes for producing compounds useful in
this invention. One general scheme (which has several
sub-variations) involves the case where both R.sub.3 and R.sub.4
are both hydrogen. This first scheme is described immediately below
in Scheme I. The other general scheme (which also has several
sub-variations) involves the case where at least one of R.sub.3 and
R.sub.4 is a moiety other than hydrogen but within the scope of
Formula I above. This second scheme is described below as "Scheme
II."
The general scheme for preparing compounds where both R.sub.3 and
R.sub.4 are both hydrogen is illustrated in Scheme I, which is
described in part in U.S. Pat. No. 3,312,730, which is incorporated
herein by reference. In Scheme I, R.sub.1 is as defined in Formula
I above. However, in Scheme I, that substituent can also be a
reactive moiety (e.g. a nitro group) that later can be reacted to
make a large number of other substituted indenes from the
nitro-substituted indenes . ##STR2##
In Scheme I, several sub-variations can be used. In one
sub-variation, a substituted benzaldehyde (a) may be condensed with
a substituted acetic ester in a Knoevenagel reaction (see reaction
2) or with an .alpha.-halogeno propionic ester in a Reformatsky
Reaction (see reactions 1 and 3). The resulting unsaturated ester
(c) is hydrogenated and hydrolyzed to give a substituted benzyl
propionic acid (e) (see reactions 4 and 5). Alternatively, a
substituted malonic ester in a typical malonic ester synthesis (see
reactions 6 and 7) and hydrolysis decarboxylation of the resulting
substituted ester (g) yields the benzyl propionic acid (e)
directly. This latter method is especially preferable for nitro and
alkylthio substituents on the benzene ring.
The next step is the ring closure of the .beta.-aryl proponic acid
(e) to form an indanone (h) which may be carried out by a
Friedel-Crafts Reaction using a Lewis acid catalyst (Cf. Organic
Reactions, Vol. 2, p. 130) or by heating with polyphosphoric acid
(see reactions 8 and 9, respectively). The indanone (h) may be
condensed with an .alpha.-halo ester in the Reformatsky Reaction to
introduce the aliphatic acid side chain by replacing the carboxyl
group (see reaction 10). Alternately, this introduction can be
carried out by the use of a Wittig Reaction in which the reagent is
a .alpha.-triphenylphosphinyl ester, a reagent which replaces the
carbonyl with a double bond to the carbon (see reaction 12). This
product (l) is then immediately rearranged into the indene (j) (see
reaction 13). If the Reformatsky Reaction route is used, the
intermediate 3-hydroxy-3-aliphatic acid derivative i must be
dehydrated to the indene (j) (see reaction 11).
The indenylacetic acid (k) in THF then is allowed to react with
oxalyl or thionyl chloride or similar reagent to produce the acid
chloride (m) (see reaction 15), whereupon the solvent is
evaporated. There are two methods to carry out reaction 16, which
is the addition of the benzylamine side chain (n).
Method (I)
In the first method, the benzylamine (n) is added slowly at room
temperature to a solution of 5-fluoro-2-methyl-3-indenylacetyl
chloride in CH.sub.2 Cl.sub.2. The reaction mixture is refluxed
overnight, and extracted with aqueous HCl (10%), water, and aqueous
NaHCO.sub.3 (5%). The organic phase is dried (Na.sub.2 SO.sub.4)
and is evaporated to give the amide compound (o)
Method (II)
In the second method, the indenylacetic acid (k) in DMA is allowed
to react with a carbodiimide (e.g.
N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride) and
benzylamine at room temperature for two days. The reaction mixture
is added dropwise to stirred ice water. A yellow precipitate is
filtered off, is washed with water, and is dried in vacuo.
Recrystallization gives the amide compound (o).
Compounds of the type a' (Scheme III), o (Scheme I), t (Scheme II),
y (Scheme IIB) may all be used in the condensation reaction shown
in Scheme III.
Substituents
X=halogen, usually Cl or Br.
E=methyl, ethyl or benzyl, or lower acyl.
R.sub.1, R.sub.2, R.sub.6, R.sub.5, and R.sub.7 =as defined in
Formula I.
Y, n and m=as defined in Formula I.
Reagents and general conditions for Scheme I (numbers refer to the
numbered reactions):
(1) Zn dust in anhydrous inert solvent such as benzene and
ether.
(2) KHSO.sub.4 or p-toluene sulfonic acid.
(3) NaOC.sub.2 H.sub.5 in anhydrous ethanol at room
temperature.
(4) H.sub.2 palladium on charcoal, 40 p.s.i. room temperature.
(5) NaOH in aqueous alcohol at 20-100.degree..
(6) NaOC.sub.2 H.sub.5 or any other strong base such as NaH or
K-t-butoxide.
(7) Acid.
(8) Friedel-Crafts Reaction using a Lewis Acid catalyst Cf. Organic
Reactions, Vol. 11, p. 130.
(9) Heat with polyphosphoric acid.
(10) Reformatsky Reaction: Zn in inert solvent, heat.
(11) p-Toluene sulfonic acid and CaCl.sub.2 or I.sub.2 at
200.degree.
(12) Wittig Reaction using (C.sub.6 H.sub.5).sub.3 P.dbd.C--COOE
20-80.degree. in ether or benzene
(13)
(a) NBS/CCl.sub.4 /benzoyl peroxide
(b) PtO.sub.2 /H.sub.2 (1 atm.)/acetic acid
(14)
(a) NaOH
(b) HCl
(15) Oxalyl or thionyl chloride in CH.sub.2 Cl.sub.2 or THF
(16)
Method I: 2 equivalents of NH.sub.2 --C(R.sub.5
R.sub.6)--Ph--(R.sub.7).
Method II: carbodiimide in THF
(17) 1N NaOCH.sub.3 in MeOH under reflux conditions
Indanones within the scope of compound (h) in Scheme I are known in
the literature and are thus readily available as intermediates for
the remainder of the synthesis so that reactions 1-7 can be
conveniently avoided. Among such known indanones are:
5-methoxyindanone
6-methoxyindanone
5-methylindanone
5-methyl-6-methoxyindanone
5-methyl-7-chloroindanone
4-methoxy-7-chloroindanone
4-isopropyl-2,7-dimethylindanone
5,6,7-trichloroindanone
2-n-butylindanone
5-methylthioindanone
Scheme II has two mutually exclusive sub-schemes: Scheme IIA and
Scheme IIB. Scheme IIA is used when R.sub.3 is hydroxy and R.sub.4
is hydrogen or when the two substituents form an oxo group. When
R.sub.3 is lower alkyl amino, Scheme IIB is employed. ##STR3##
Similar to Scheme I, in Scheme IIA the indenylacetic acid (k) in
THF is allowed to react with oxalylchloride under reflux conditions
to produce the acid chloride (p) (see reaction 18), whereupon the
solvent is evaporated. In reaction 19, a 0.degree. C. mixture of a
benzyl hydroxylamine hydrochloride (q) and Et.sub.3 N is treated
with a cold solution of the acid chloride in CH.sub.2 Cl.sub.2 over
a period of 45-60 minutes. The mixture is warmed to room
temperature and stirred for one hour, and is treated with water.
The resulting organic layer is washed with 1 N HCl and brine, is
dried over magnesium sulfate and is evaporated. The crude product,
a N-hydroxy-N-benzyl acetamide (r) is purified by crystallization
or flash chromatography. This general procedure is taught by
Hoffman et al., JOC 1992, 57, 5700-5707.
The next step is the preparation of the N-mesyloxy amide (s) in
reaction 20, which is also taught by Hoffman et al., JOC 1992, 57,
5700-5707. Specifically, to a solution of the hydroxamic acid (r)
in CH.sub.2 Cl.sub.2 at 0.degree. C. is added triethylamine. The
mixture is stirred for 10-12 minutes, and methanesulfonyl chloride
is added dropwise. The mixture is stirred at 0.degree. C. for two
hours, is allowed to warm to room temperature, and is stirred for
another two hours. The organic layer is washed with water, 1 N HCl,
and brine, and is dried over magnesium sulfate. After rotary
evaporation, the product(s) is usually purified by crystallization
or flash chromatography.
The preparation of the N-benzyl-.alpha.-(hydroxy) amide (t) in
reaction 21, is also taught by Hoffman et al., JOC 1992, 57,
5700-5707 and Hoffman et al., JOC 1995, 60, 4121-4125.
Specifically, to a solution of the N-(mesyloxy) amide (s) in
CH.sub.3 CN/H.sub.2 O is added triethylamine in CH.sub.3 CN over a
period of 6-12 hours. The mixture is stirred overnight. The solvent
is removed, and the residue is dissolved in ethyl acetate. The
solution is washed with water, 1 N HCl, and brine, and is dried
over magnesium sulfate. After rotary evaporation, the product (t)
is usually purified by recrystallization.
Reaction 22 in Scheme IIA involves a condensation with certain
aldehydes, which is described in Scheme III below, a scheme that is
common to products made in accordance with Schemes I, IIA and
IIB.
The final reaction 23 in Scheme IIA is the preparation of the
N-benzyl-.alpha.-ketoamide (v), which involves the oxidation of a
secondary alcohol (u) to a ketone by e.g. a Pfitzner-Moffatt
oxidation, which selectively oxidizes the alcohol without oxidizing
the Y group. Compounds (u) and (v) may be derivatized in order to
obtain compounds with R.sub.3 and R.sub.4 groups as set forth in
Formula I. ##STR4##
As explained above, Scheme IIB is employed when R.sub.3 is lower
alkyl amino. Similar to Scheme I, in Scheme IIB the indenylacetic
acid (k) in THF is allowed to react with oxalylchloride under
reflux conditions to produce the acid chloride (p) (see reaction
18), whereupon the solvent is evaporated. In reaction 24, a mixture
of an alkyl hydroxylamine hydrochloride (i.e. HO--NHR where R is a
lower alkyl, preferably isopropyl) and Et.sub.3 N is treated at
0.degree. C. with a cold solution of the acid chloride in CH.sub.2
Cl.sub.2 over a period of 45-60 minutes. The mixture is warmed to
room temperature and is stirred for one hour, and is diluted with
water. The resulting organic layer is washed with 1 N HCl and
brine, is dried over magnesium sulfate and is evaporated. The crude
product, a N-hydroxy-N-alkyl acetamide (w) is purified by
crystallization or flash chromatography. This general procedure is
also taught by Hoffman et al., JOC 1992, 57, 5700-5707.
The preparation of the N-mesyloxy amide (x) in reaction 25, which
is also taught by Hoffman et al., JOC 1992, 57, 5700-5707.
Specifically, a solution of the hydroxamic acid (w) in CH.sub.2
Cl.sub.2 at 0.degree. C. is treated with triethylamine, is stirred
for 10-12 minutes, and is treated dropwise with methanesulfonyl
chloride. The mixture is stirred at 0.degree. C. for two hours, is
allowed to warm to room temperature, and is stirred for another two
hours. The resulting organic layer is washed with water, 1 N HCl,
and brine, and is dried over magnesium sulfate. After rotary
evaporation, the product (x) is usually purified by crystallization
or flash chromatography.
The preparation of the N-benzyl
indenyl-.alpha.-loweralkylamino-acetamide compound (y) in Scheme
IIB as taught by Hoffman et al., JOC 1995, 60, 4121-25 and J. Am.
Chem Soc. 1993, 115, 5031-34, involves the reaction of the
N-mesyloxy amide (x), with a benzylamine in CH.sub.2 Cl.sub.2 at
0.degree. C. is added over a period of 30 minutes. The resulting
solution is stirred at 0.degree. C. for one hour and at room
temperature overnight. The solvent is removed, and the residue is
treated with 1 N NaOH. The extract with CH.sub.2 Cl.sub.2 is washed
with water and is dried over magnesium sulfate. After rotary
evaporation, the product (y) is purified by flash chromatography or
crystallization. ##STR5##
Scheme III involves the condensation of the heterocycloaldehydes
(i.e. Y--CHO) with the indenyl amides to produce the final
compounds of Formula I. This condensation is employed, for example,
in reaction 17 in Scheme I above and in reaction 22 in Scheme IIA.
It is also used to convert compound (y) in Scheme IIB to final
compounds of Formula I.
In Scheme III, the amide (a') from the above schemes, a
N-heterocycloaldehyde (z), and sodium methoxide (1 M in methanol)
are stirred at 60.degree. C. under nitrogen for 24 hours. After
cooling, the reaction mixture is poured into ice water. A solid is
filtered off, is washed with water, and is dried in vacuo.
Recrystallization provides a compound of Formula I in Schemes I and
IIB and the intermediate (u) in Scheme IIA.
As has been pointed out above, it is preferable in the preparation
of many types of the compounds of this invention, to use a nitro
substituent on the benzene ring of the indanone nucleus and convert
it later to a desired substituent since by this route a great many
substituents can be reached. This is done by reduction of the nitro
to the amino group followed by use of the Sandmeyer Reaction to
introduce chlorine, bromine, cyano or xanthate in place of the
amino. From the cyano derivatives hydrolysis yields the carboxamide
and carboxylic acid; other derivatives of the carboxy group such as
the esters can then be prepared. The xanthates, by hydrolysis,
yield the mercapto group that may be oxidized readily to the
sulfonic acid or alkylated to an alkylthio group which can then be
oxidized to alkylsulfonyl groups. These reactions may be carried
out either before or after the introduction of the
1-substituent.
The foregoing may be better understood from the following examples
that are presented for purposes of illustration and are not
intended to limit the scope of the invention. As used in the
following examples, the references to substituents such as R.sub.1,
R.sub.2, etc., refer to the corresponding compounds and
substituents in Formula I above.
EXAMPLE 1
(Z)-5-Fluoro-2-Methyl-(4-Pyridinylidene)-3-(N-Benzyl)-Indenylacetamide
(A) p-Fluoro-.alpha.-methylcinnamic acid
p-Fluorobenzaldehyde (200 g, 1.61 mol), propionic anhydride (3.5 g,
2.42 mol) and sodium propionate (155 g, 1.61 mol) are mixed in a
one liter three-necked flask which had been flushed with nitrogen.
The flask is heated gradually in an oil-bath to 140.degree. C.
After 20 hours, the flask is cooled to 100.degree. C. and poured
into 8 l of water. The precipitate is dissolved by adding potassium
hydroxide (302 g) in 2 l of water. The aqueous solution is
extracted with ether, and the ether extracts are washed with
potassium hydroxide solution. The combined aqueous layers are
filtered, are acidified with concentrated HCl, and are filtered.
The collected solid, p-fluoro-.alpha.-methylcinnamic acid, is
washed with water, and is dried and used as obtained.
(B) p-Fluoro-.alpha.-methylhydrocinnamic acid
To p-fluoro-.alpha.-methylcinnamic acid (177.9 g, 0.987 mol) in 3.6
l ethanol is added 11.0 g of 5% Pd/C. The mixture is reduced at
room temperature under a hydrogen pressure of 40 p.s.i. When
hydrogen uptake ceases, the catalyst is filtered off, and the
solvent is evaporated in vacuo to give the product,
p-fluoro-.alpha.-methylhydrocinnamic acid, which was used directly
in the next step.
(C) 6-Fluoro-2-methylindanone
To 932 g polyphosphoric acid at 70.degree. C. (steam bath) is added
p-fluoro-.alpha.-methylhydrocinnamic acid (93.2 g, 0.5 mol) slowly
with stirring. The temperature is gradually raised to 95.degree.
C., and the mixture is kept at this temperature for 1 hour. The
mixture is allowed to cool and is added to 2 l. of water. The
aqueous suspension is extracted with ether. The extract is washed
twice with saturated sodium chloride solution, 5% Na.sub.2 CO.sub.3
solution, and water, and is dried, and is concentrated on 200 g
silica-gel; the slurry is added to a five pound silica-gel column
packed with 5% ether-petroleum ether. The column is eluted with
5-10% ether-petroleum ether, to give 6-fluoro-2-methylindanone.
Elution is followed by TLC.
(D) 5-fluoro-2-methylindenyl-3-acetic acid
A mixture of 6-fluoro-2-methylindanone (18.4 g, 0.112 mol),
cyanoacetic acid (10.5 g, 0.123 mol), acetic acid (6.6 g), and
ammonium acetate (1.7 g) in dry toluene (15.5 ml) is refluxed with
stirring for 21 hours, as the liberated water is collected in a
Dean Stark trap. The toluene is evaporated, and the residue is
dissolved in 60 ml of hot ethanol and 14 ml of 2.2 N aqueous
potassium hydroxide solution. 22 g of 85% KOH in 150 ml of water is
added, and the mixture refluxed for 13 hours under nitrogen. The
ethanol is removed under vacuum, and 500 ml water is added. The
aqueous solution is extracted well with ether, and is then boiled
with charcoal. The aqueous filtrate is acidified to pH 2 with 50%
cold hydrochloric acid. The precipitate is dried and
5-fluoro-2-methylindenyl-3-acetic acid (M.P. 164-166.degree. C.) is
obtained.
(E) 5-fluoro-2-methylindenyl-3-acetyl chloride
5-fluoro-2-methylindenyl-3-acetic acid (70 mmol) in THF (70 ml) is
allowed to react with oxalylchloride (2 M in CH.sub.2 Cl.sub.2 ; 35
ml; 70 mmol) under reflux conditions (24 hours). The solvent is
evaporated to yield the title compound, which is used as such in
the next step.
(F) 5-Fluoro-2-methyl-3-(N-benzyl)-indenylacetamide
Benzylamine (5 mmol) is added slowly at room temperature to a
solution of 5-fluoro-2-methylindenyl-3-acetyl chloride (2.5 mmol.)
in CH.sub.2 Cl.sub.2 (10 ml). The reaction mixture is refluxed
overnight, and is extracted with aqueous HCl (10%), water, and
aqueous NaHCO.sub.3 (5%). The organic phase is dried (Na.sub.2
SO.sub.4) and is evaporated to give the title compound, which is
recrystallized from CH.sub.2 Cl.sub.2 to give the title compound as
a white solid (m.p. 144.degree. C.).
(G)
(Z)-5-Fluoro-2-methyl-(4-pyridinylidene)-3-(N-benzyl)-indenylacetamide
5-fluoro-2-methyl-3-(N-benzyl)-indenylacetamide (3.38 mmol),
4-pyridinecarboxaldehyde (4 mmol), sodium methoxide (1M NaOCH.sub.3
in methanol (30 ml)) are heated at 60.degree. C. under nitrogen
with stirring for 24 hours. After cooling, the reaction mixture is
poured into ice water (200 ml). A solid is filtered off, washed
with water, and dried in vacuo. Recrystallization from CH.sub.3 CN
gives the title compound (m.p. 202.degree. C.) as a yellow solid
(R.sub.1 =F, R.sub.2 =CH.sub.3, R.sub.3 =H, R.sub.4 =H, R.sub.5 =H,
R.sub.6 =H, R.sub.7 =H, n=1, m=1, Y=4-pyridinyl).
(H)
(E)-5-Fluoro-2-methyl-(4-pyridinylidene)-3-(N-benzyl)-indenylacetamide
The mother liquor obtained from the CH.sub.3 CN recrystallization
of 1G is rich on the geometrical isomer of 1G. The E-isomer can be
obtained pure by repeated recrystallizations from CH.sub.3 CN.
EXAMPLE 2
(Z)-5-Fluoro-2-Methyl-(3-Pyridinylidene)-3-(N-Benzyl)-Indenylacetamide
This compound is obtained from
5-fluoro-2-methyl-3-(N-benzyl)-indenylacetamide (Example 1F) using
the procedure of Example 1, part G and replacing
4-pyridinecarboxaldehyde with 3-pyridinecarboxaldehyde.
Recrystallization from CH.sub.3 CN gives the title compound (m.p.
175.degree. C.)(R.sub.1 =F, R.sub.2 =CH.sub.3, R.sub.3 =H, R.sub.4
=H, R.sub.5 =H, R.sub.6 =H, R.sub.7 =H, n=1, m=1,
Y=3-pyridinyl).
EXAMPLE 3
(Z)-5-Fluoro-2-Methyl-(2-Pyridinylidene)-3-(N-Benzyl)-Indenylacetamide
This compound is obtained from
5-fluoro-2-methyl-3-(N-benzyl)-indenylacetamide (Example 1F) using
the procedure of Example 1, part G and replacing
4-pyridinecarboxaldehyde with 2-pyridinecarboxaldehyde.
Recrystallization from ethylacetate gives the title compound (m.p.
218.degree. C.)(R.sub.1 =F, R.sub.2 =CH.sub.3, R.sub.3 =H, R.sub.4
=H, R.sub.5 =H, R.sub.6 =H, R.sub.7 =H, n=1, m=1,
Y=2-pyridinyl).
EXAMPLE 4
(Z)-5-Fluoro-2-Methyl-(4-Quinolinylidene)-3-(N-Benzyl)-Indenylacetamide
This compound is obtained from
5-fluoro-2-methyl-3-(N-benzyl)-indenylacetamide (Example 1F) using
the procedure of Example 1, part G and replacing
4-pyridinecarboxaldehyde with 4-quinolinecarboxaldehyde.
Recrystallization from ethylacetate gives the title compound (m.p.
239.degree. C.)(R.sub.1 =F, R.sub.2 =CH.sub.3, R.sub.3 =H, R.sub.4
=H, R.sub.5 =H, R.sub.6 =H, R.sub.7 =H, n=1, m=1,
Y=4-quinolinyl).
EXAMPLE 5
(Z)-5-Fluoro-2-Methyl-(4,6-Dimethyl-2-Pyridinylidene)-3-(N-Benzyl)-Indenyla
cetamide
5-Fluoro-2-methyl-3-(N-benzyl)-indenylacetamide from Example 1,
part F is allowed to react with
4,6-dimethyl-2-pyridinecarboxaldehyde according to the procedure of
Example 1, part G in order to obtain the title compound.
Recrystallization gives the title compound (R.sub.1 =F, R.sub.2
=CH.sub.3, R.sub.3 =H, R.sub.4 =H, R.sub.5 =H, R.sub.6 =H, R.sub.7
=H, n=1, m=1, Y=4,6-dimethyl-2-pyridinyl).
EXAMPLE 6
(Z)-5-Fluoro-2-Methyl-(3-Quinolinylidene)-3-(N-Benzyl)-Indenylacetamide
5-Fluoro-2-methyl-3-(N-benzyl)-indenylacetamide from Example 1,
part F is allowed to react with 3-quinolinecarboxaldehyde according
to the procedure of Example 1, part G in order to obtain the title
compound. Recrystallization gives the title compound (R.sub.1 =F,
R.sub.2 =CH.sub.3, R.sub.3 =H, R.sub.4 =H, R.sub.5 =H, R.sub.6 =H,
R.sub.7 =H, n=1, m=1, Y=3-quinolinyl)
EXAMPLE 7
(Z)-5-Fluoro-2-Methyl-(2-Quinolinylidene)-3-(N-Benzyl)-Indenylacetamide
5-Fluoro-2-methyl-3-(N-benzyl)-indenylacetamide from Example 1,
part F is allowed to react with 2-quinolinecarboxaldehyde according
to the procedure of Example 1, part G in order to obtain the title
compound. Recrystallization gives the title compound (R.sub.1 =F,
R.sub.2 =CH.sub.3, R.sub.3 =H, R.sub.4 =H, R.sub.5 =H, R.sub.6 =H,
R.sub.7 =H, n=1, m=1, Y=2-quinolinyl).
EXAMPLE 8
(Z)-5-Fluoro-2-Methyl-(Pyrazinylidene)-3-(N-Benzyl)-Indenylacetamide
5-Fluoro-2-methyl-3-(N-benzyl)-indenylacetamide from Example 1,
part F is allowed to react with pyrazinealdehyde according to the
procedure of Example 1, part G in order to obtain the title
compound. Recrystallization gives the title compound (R.sub.1 =F,
R.sub.2 =CH.sub.3, R.sub.3 =H, R.sub.4 =H, R.sub.5 =H, R.sub.6 =H,
R.sub.7 =H, n=1,m=1, Y=pyrazinyl).
EXAMPLE 9
(Z)-5-Fluoro-2-Methyl-(3-Pydidazinylidene)-3-(N-Benzyl)-Indenylacetamide
5-Fluoro-2-methyl-3-(N-benzyl)-indenylacetamide from Example 1,
part F is allowed to react with pyridazine-3-aldehyde according to
the procedure of Example 1, part G in order to obtain the title
compound. Recrystallization gives the title compound (R.sub.1 =F,
R.sub.2 =CH.sub.3, R.sub.3 =H, R.sub.4 =H, R.sub.5 =H, R.sub.6 =H,
R.sub.7 =H, n=1, m=1, Y=3-pyridazinyl).
EXAMPLE 10
(Z)-5-Fluoro-2-Methyl-(4-Pyrimidinylidene)-3-(N-Benzyl)-Indenylacetamide
5-Fluoro-2-methyl-3-(N-benzyl)-indenylacetamide from Example 1,
part F is allowed to react with pyrimidine-4-aldehyde according to
the procedure of Example 1, part G in order to obtain the title
compound. Recrystallization gives the title compound (R.sub.1 =F,
R.sub.2 =CH.sub.3, R.sub.3 =H, R.sub.4 =H, R.sub.5 =H, R.sub.6 =H,
R.sub.7 =H, n=1, m=1, Y=4-pyrimidinyl).
EXAMPLE 11
(Z)-5-Fluoro-2-Methyl-(2-Methyl-4-Pyrimidinylidene)-3-(N-Benzyl)-Indenylace
tamide
5-Fluoro-2-methyl-3-(N-benzyl)-indenylacetamide from Example 1,
part F is allowed to react with 2-methyl-pyrimidine-4-aldehyde
according to the procedure of Example 1, part G in order to obtain
the title compound. Recrystallization gives the title compound
(R.sub.1 =F, R.sub.2 =CH.sub.3, R.sub.3 =H, R.sub.4 =H, R.sub.5 =H,
R.sub.6 =H, R.sub.7 =H, n=1, m=1, Y=2-methyl-4-pyrimidinyl).
EXAMPLE 12
(Z)-5-Fluoro-2-Methyl-(4-Pyridazinylidene)-3-(N-Benzyl)-Indenylacetamide
5-Fluoro-2-methyl-3-(N-benzyl)-indenylacetamide from Example 1,
part F is allowed to react with pyridazine-4-aldehyde according to
the procedure of Example 1, part G in order to obtain the title
compound. Recrystallization gives the title compound (R.sub.1 =F,
R.sub.2 =CH.sub.3, R.sub.3 =H, R.sub.4 =H, R.sub.5 =H, R.sub.6 =H,
R.sub.7 =H, n=1, m=, Y=4-pyridazinyl).
EXAMPLE 13
(Z)-5-Fluoro-2-Methyl-(1-Methyl-3-Indolylidene)-3-(N-Benzyl)-Indenylacetami
de
5-Fluoro-2-methyl-3-(N-benzyl)-indenylacetamide from Example 1,
part F is allowed to react with 1-methylindole-3-carboxaldehyde
according to the procedure of Example 1, part G in order to obtain
the title compound. Recrystallization gives the title compound
(R.sub.1 =F, R.sub.2 =CH.sub.3, R.sub.3 =H, R.sub.4 =H, R.sub.5 =H,
R.sub.6 =H, R.sub.7 =H, n=1, m=1, Y=1-methyl-3-indolyl).
EXAMPLE 14
(Z)-5-Fluoro-2-Methyl-(1-Acetyl-3-Indolylidene)-3-(N-Benzyl)-Indenylacetami
de
5-Fluoro-2-methyl-3-(N-benzyl)-indenylacetamide from Example 1,
part F is allowed to react with 1-acetyl-3-indolecarboxaldehyde
according to the procedure of Example 1, part G in order to obtain
the title compound. Recrystallization gives the title compound
(R.sub.1 =F, R.sub.2 =CH.sub.3, R.sub.3 =H, R.sub.4 =H, R.sub.5 =H,
R.sub.6 =H, R.sub.7 =H, n=1, m=1, Y=1-acetyl-3-indolyl).
EXAMPLE 15
(Z)-5-Fluoro-2-Methyl-(4-Pyridinylidene)-3-(N-2-fluorobenzyl)-Indenylacetam
ide
(A) 5-Fluoro-2-methyl-3-(N-2-fluorobenzyl)-indenylacetamide
This compound is obtained from 5-fluoro-2-methylindenyl-3-acetyl
chloride (Example 1E) using the procedure of Example 1, Part F and
replacing benzylamine with 2-fluorobenzylamine.
(B)
(Z)-5-Fluoro-2-methyl-(4-pyridinylidene)-3-(N-2-fluorobenzyl-indenylacetam
ide
5-Fluoro-2-methyl-3-(N-2-fluorobenzyl)-indenylacetamide is allowed
to react with 4-pryidinecarboxaldehyde according to the procedure
of Example 1, part G in order to obtain the title compound.
Recrystallization gives the title compound (R.sub.1 =F, R.sub.2
=CH.sub.3, R.sub.3 =H, R.sub.4 =H, R.sub.5 =H, R.sub.6 =H, R.sub.7
=F, n=1, m=1, Y=4-pyridinyl).
EXAMPLE 16
(Z)-5-Fluoro-2-Methyl-(3-Pyridinylidene)-3-(N-2-Fluorobenzyl)-Indenylacetam
ide
5-Fluoro-2-methyl-3-(N-2-fluorobenzyl)-indenylacetamide from
Example 15, part A is allowed to react with
3-pryidinecarboxaldehyde according to the procedure of Example 1,
part G in order to obtain the title compound. Recrystallization
gives the title compound (R.sub.1 =F, R.sub.2 =CH.sub.3, R.sub.3
=H, R.sub.4 =H, R.sub.5 =H, R.sub.6 =H, R.sub.7 =F, n=1, m=1,
Y=3-pyridinyl).
EXAMPLE 17
(Z)-5-Fluoro-2-Methyl-(2-Pyridinylidene)-3-(N-2-Fluorobenzyl)-Indenylacetam
ide
5-Fluoro-2-methyl-3-(N-2-fluorobenzyl)-indenylacetamide from
Example 15, part A is allowed to react with
2-pyridinecarboxaldehyde according to the procedure of Example 1,
part G in order to obtain the title compound. Recrystallization
gives the title compound (R.sub.1 =F, R.sub.2 =CH.sub.3, R.sub.3
=H, R.sub.4 =H, R.sub.5 =H, R.sub.6 =H, R.sub.7 =F, n=1, m=1,
Y=2-pyridinyl).
EXAMPLE 18
(Z)-5-Fluoro-2-Methyl-(4-Quinolinylidene)-3-(N-2-Fluorobenzyl)-Indenylaceta
mide
5-Fluoro-2-methyl-3-(N-2-fluorobenzyl)-indenylacetamide from
Example 15, part A is allowed to react with
4-quinolinecarboxaldehyde according to the procedure of Example 1,
part G in order to obtain the title compound. Recrystallization
gives the title compound (R.sub.1 =F, R.sub.2 =CH.sub.3, R.sub.3
=H, R.sub.4 =H, R.sub.5 =H, R.sub.6 =H, R.sub.7 =F, n=1, m=1,
Y=3-quinolinyl).
EXAMPLE 19
(Z)-5-Fluoro-2-Methyl-(3-Pyrazinylidene)-3-(N-2-Fluorobenzyl)-Indenylacetam
ide
5-Fluoro-2-methyl-3-(N-2-fluorobenzyl)-indenylacetamide from
Example 15, part A is allowed to react with pyrazinealdehyde
according to the procedure of Example 1, Part G in order to obtain
the title compound. Recrystallization gives the title compound
(R.sub.1 =F, R.sub.2 =CH.sub.3, R.sub.3 =H, R.sub.4 =H, R.sub.5 =H,
R.sub.6 =H, R.sub.7 =F, n=1, m=1, Y=3-pyrazinyl).
EXAMPLE 20
(Z)-5-Fluoro-2-Methyl-(3-Pyridazinylidene)-3-(N-2-Fluorobenzyl)-Indenylacet
amide
5-Fluoro-2-methyl-3-(N-2-fluorobenzyl)-indenylacetamide from
Example 15, part A is allowed to react with
3-pryidaziine-3-aldehyde according to the procedure of Example 1,
Part G in order to obtain the title compound. Recrystallization
gives the title compound (R.sub.1 =F, R.sub.2 =CH.sub.3, R.sub.3
=H, R.sub.4 =H, R.sub.5 =H, R.sub.6 =H, R.sub.7 =F, n=1, m=1,
Y=3-pyridazinyl).
EXAMPLE 21
(Z)-5-Fluoro-2-Methyl-(3-Pyrimidinylidene)-3-(N-2-Fluorobenzyl)-Indenylacet
amide
5-Fluoro-2-methyl-3-(N-2-fluorobenzyl)-indenylacetamide from
Example 15, part A is allowed to react with pryimidine-4-aldehyde
according to the procedure of Example 1, Part G in order to obtain
the title compound. Recrystallization gives the title compound
(R.sub.1 =F, R.sub.2 =CH.sub.3, R.sub.3 =H, R.sub.4 =H, R.sub.5 =H,
R.sub.6 =H, R.sub.7 =F, n=1, m=1, Y=3-pyrimidinyl).
EXAMPLE 22
(Z)-5-Fluoro-2-Methyl-(4-Pyrdazinylidene)-3-(N-2-Fluorobenzyl)-Indenylaceta
mide
5-Fluoro-2-methyl-3-(N-2-fluorobenzyl)-indenylacetamide from
Example 15, part A is allowed to react with pryidazine-4-aldehyde
according to the procedure of Example 1, Part G in order to obtain
the title compound. Recrystallization gives the title compound
(R.sub.1 =F, R.sub.2 =CH.sub.3, R.sub.3 =H, R.sub.4 =H, R.sub.5 =H,
R.sub.6 =H, R.sub.7 =F, n=1, m=1, Y=4-pyridazinyl).
EXAMPLE 23
(Z)-5-Fluoro-2-Methyl-(4-Pyridinylidene)-3-(N-(S-.alpha.-Hydroxymethyl)Benz
yl)-Indenylacetamide
(A)
5-Fluoro-2-methyl-3-(N-(S-.alpha.-hydroxylmethyl)benzyl)-indenylacetamide
5-Fluoro-2-methylindenyl-3-acetic acid (from Example 1D) (2.6 mmol)
in DMA (2 ml) is allowed to react with
n-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride (4
mmol) and S-2-amino-2-phenylethanol (3.5 mmol) at room temperature
for two days. The reaction mixture is added dropwise to stirred ice
water (50 ml). A white precipitate is filtered off, washed with
water (5 ml), and dried in vacuo. Recrystallization from
ethylacetate gives the desired compound.
(B)
(Z)-5-fluoro-2-methyl-(4-pyridinylidene)-3-(N-(S-.alpha.-hydroxymethyl)ben
zyl)-indenylacetamide
5-Fluoro-2-methyl-3-(N-(S-.alpha.-hydroxylmethyl)benzyl)-indenylacetamide
from part A is allowed to react with 4-pryidinecarboxaldehyde
according to the procedure of Example 1, Part G in order to obtain
the title compound. Recrystallization gives the title compound
(R.sub.1 =F, R.sub.2 =CH.sub.3, R.sub.3 =H, R.sub.4 =H, R.sub.5
=CH.sub.2 OH, R.sub.6 =H, R.sub.7 =H, n=1, m=1, Y=4-pyridinyl).
EXAMPLE 24
(Z)-5-Fluoro-2-Methyl-(3-Pyridinylidene)-3-(N-(S-.alpha.-Hydroxymethyl)Benz
yl)-Indenylacetamide
5-Fluoro-2-methyl-3-(N-(S-.alpha.-hydroxylmethyl)benzyl)-indenylacetamide
from Example 23 part A is allowed to react with
3-pryidinecarboxaldehyde according to the procedure of Example 1,
Part G in order to obtain the title compound. Recrystallization
gives the title compound (R.sub.1 =F, R.sub.2 =CH.sub.3, R.sub.3
=H, R.sub.4 =H, R.sub.5 =CH.sub.2 OH, R.sub.6 =H, R.sub.7 =H, n=1,
m=1, Y=3-pyridinyl).
EXAMPLE 25
(Z)-5-Fluoro-2-Methyl-(2-Pyridinylidene)-3-(N-(S-.alpha.-Hydroxymethyl)Benz
yl)-Indenylacetamide
5-Fluoro-2-methyl-3-(N-(S-.alpha.-hydroxylmethyl)benzyl)-indenylacetamide
from Example 23 part A is allowed to react with
2-pryidinecarboxaldehyde according to the procedure of Example 1,
Part G in order to obtain the title compound. Recrystallization
gives the title compound (R.sub.1 =F, R.sub.2 =CH.sub.3, R.sub.3
=H, R.sub.4 =H, R.sub.5 =CH.sub.2 OH, R.sub.6 =H, R.sub.7 =H, n=1,
m=1, Y=2-pyridinyl).
EXAMPLE 26
(Z)-5-Fluoro-2-Methyl-(4-Quinolinylidene)-3-(N-(S-.alpha.-Hydroxymethyl)Ben
zyl-Indenylacetamide
5-Fluoro-2-methyl-3-(N-(S-.alpha.-hydroxylmethyl)benzyl)-indenylacetamide
from Example 23 part A is allowed to react with
4-quinolinecarboxaldehyde according to the procedure of Example 1,
Part G in order to obtain the title compound. Recrystallization
gives the title compound (R.sub.1 =F, R.sub.2 =CH.sub.3, R.sub.3
=H, R.sub.4 =H, R.sub.5 =CH.sub.2 OH, R.sub.6 =H, R.sub.7 =H, n=1,
m=1, Y=4-quinolinyl).
EXAMPLE 27
(Z)-5-Fluoro-2-Methyl-(Pyrazidinylidene)-3-(N-(S-.alpha.-Hydroxymethyl)Benz
yl)-Indenylacetamide
5-Fluoro-2-methyl-3-(N-(S-.alpha.-hydroxylmethyl)benzyl)-indenylacetamide
from Example 23 part A is allowed to react with
pryazidinecarboxaldehyde according to the procedure of Example 1,
Part G in order to obtain the title compound. Recrystallization
gives the title compound (R.sub.1 =F, R.sub.2 =CH.sub.3, R.sub.3
=H, R.sub.4 =H, R.sub.5 =CH.sub.2 OH, R.sub.6 =H, R.sub.7 =H, n=1,
m=1, Y=pyrazidinyl).
EXAMPLE 28
(Z)-5-Fluoro-2-Methyl-(3-Pyridazinylidene)-3-(N-(S-.alpha.-Hydroxymethyl)Be
nzyl)-Indenylacetamide
5-Fluoro-2-methyl-3-(N-(S-.alpha.-hydroxylmethyl)benzyl)-indenylacetamide
from Example 23 part A is allowed to react with
pryidazine-3-aldehyde according to the procedure of Example 1, Part
G in order to obtain the title compound. Recrystallization gives
the title compound (R.sub.1 =F, R.sub.2 =CH.sub.3, R.sub.3 =H,
R.sub.4 =H, R.sub.5 =CH.sub.2 OH, R.sub.6 =H, R.sub.7 =H, n=1, m=1,
Y=3-pyridazinyl).
EXAMPLE 29
(Z)-5-Fluoro-2-Methyl-(4-Pyrimidinylidene)-3-(N-(S-.alpha.-Hydroxymethyl)Be
nzyl)-Indenylacetamide
5-Fluoro-2-methyl-3-(N-(S-.alpha.-hydroxylmethyl)benzyl)-indenylacetamide
from Example 23 part A is allowed to react with
pryimidine-4-aldehyde according to the procedure of Example 1, Part
G in order to obtain the title compound. Recrystallization gives
the title compound (R.sub.1 =F, R.sub.2 =CH.sub.3, R.sub.3 =H,
R.sub.4 =H, R.sub.5 =CH.sub.2 OH, R.sub.6 =H, R.sub.7 =H, n=1, m=1,
Y=4-pyrimidinyl).
EXAMPLE 30
(Z)-5-Fluoro-2-Methyl-(4-Pydidazinylidene)-3-(N-(S-.alpha.-Hydroxymethyl)Be
nzyl)-Indenylacetamide
5-Fluoro-2-methyl-3-(N-(S-.alpha.-hydroxylmethyl)benzyl)-indenylacetamide
from Example 23 part A is allowed to react with
pryidazine-4-aldehyde according to the procedure of Example 1, Part
G in order to obtain the title compound. Recrystallization gives
the title compound (R.sub.1 =F, R.sub.2 =CH.sub.3, R.sub.3 =H,
R.sub.4 =H, R.sub.5 =CH.sub.2 OH, R.sub.6 =H, R.sub.7 =H, n=1, m=1,
Y=4-pyridazinyl).
EXAMPLE 31
rac-(Z)-5-Fluoro-2-Methyl-(4-Pyridinylidene)-3-(N-Benzyl)Indenyl-.alpha.-Hy
droxyacetamide
(A) 5-fluoro-2-methyl-3-(N-benzyl-N-hydroxy)-indenylacetamide
To a mixture of N-benzylhydroxylamine hydrochoride (12 mmol) and
Et.sub.3 N (22 mmol) in CH.sub.2 Cl.sub.2 (100 ml) at 0.degree. C.
is added a cold solution of 5-fluoro-2-methylindenyl-3-acetyl
chloride (Example 1, Step E) (10 mmol) in CH.sub.2 Cl.sub.2 (75 ml)
over a period of 45-60 minutes. The mixture is warmed to room
temperature and is stirred for 1 hour. The mixture is diluted with
water (100 ml), and the organic layer is washed with HCl
(2.times.25 ml) and brine (2.times.100 ml), dried (MgSO.sub.4) and
evaporated. The crude product is purified with flash chromatography
to give the title compound.
(B) 5-Fluoro-2-methyl-3-(N-benzyl-N-mesyloxy)-indenylacetamide
To a solution of
5-fluoro-2-methyl-3-(N-benzyl-N-hydroxy)-indenylacetamide (5 mmol)
in CH.sub.2 Cl.sub.2 (25 ml) at 0.degree. C. is added triethylamine
(5 mmol). The mixture is stirred for 10 minutes, and
methanesulfonyl chloride (5.5 mmol) is added dropwise. The solution
is stirred at 0.degree. C. for 2 hours, allowed to warm to room
temperature, and stirred for another 2 hours. The organic layer is
washed with water (2.times.20 ml), in HCl (15 ml), and brine (20
ml) and dried over MgSO.sub.4. After rotary evaporation, the
product is purified with flash chromatography to give the title
compound.
(C)
rac-5-Fluoro-2-methyl-3-(N-benzyl)-.alpha.-hydroxyindenylacetamide
To a solution of
5-fluoro-2-methyl-3-(N-benzyl-N-mesyloxy)-indenylacetamide (2 mmol)
in CH.sub.3 CN/H.sub.2 O (12 ml. each) is added triethylamine (2.1
mmol) in CH.sub.3 CN (24 ml) over a period of 6 hours. The mixture
is stirred overnight. The solvent is removed, and the residue
diluted with ethyl acetate (60 ml), washed with water (4.times.20
ml), in HCl (15 ml), and brine (20 ml) and dried over MgSO.sub.4.
After rotary evaporation, the product is purified by
recrystallization to give the title compound.
(D)
rac-(Z)-5-Fluoro-2-methyl-(4-pyridinylidene)-3-(N-benzyl)-indenyl-.alpha.-
hydroxyacetamide is obtained from
rac-5-fluoro-2-methyl-3-(N-benzyl)-.alpha.-hydroxyindenylacetamide
using the procedure of Example 1, Part G (R.sub.1 =F, R.sub.2
=CH.sub.3, R.sub.3 =OH, R.sub.4 =H, R.sub.5 =H, R.sub.6 =H, R.sub.7
=H, n=1, m=1, Y=4-pyridinyl).
EXAMPLE 32
2-[(Z)-5-Fluoro-2-Methyl-(4-Pyridinylidene)-3-(N-Benzyl)-Indenyl]-Oxyacetam
ide
For Pfitzner-Moffatt oxidation, a solution of
rac-(Z)-5-fluoro-2-methyl-(4-pyridinylidene)-3-(N-benzyl)-indenyl-.alpha.-
hydroxyacetamide (1 mmol) in DMSO (5 ml) is treated with
dicyclohexylcarbodiimide (3 mmol). The mixture is stirred
overnight, and the solvent is evaporated. The crude product is
purified by flash chromatography to give the title compound
(R.sub.1 =F, R.sub.2 =CH.sub.3, R.sub.3 and R.sub.4 together form
C.dbd.O, R.sub.5 =H, R.sub.6 =H, R.sub.7 =H, n=1, m=1, and
Y=4-pyridinyl).
EXAMPLE 33
rac-(Z)-5-Fluoro-2-Methyl-(4-Pyridinylidene)-3-(N-Benzyl)-Indenyl-.alpha.-(
2-Propylamino)-Acetamide
(A) 5-Fluoro-2-methyl-3-(N-2-propyl-N-hydroxy)-indenylacetamide is
obtained from 5-fluoro-2-methylindenyl-3-acetyl chloride (Example
1, Step E) using the procedure of Example 31, Part A and replacing
N-benzylhydroxylamine hydrochloride with N-2-propyl hydroxylamine
hydrochloride.
(B) 5-Fluoro-2-methyl-3-(N-2-propyl-N-mesyloxy)-indenylacetamide is
obtained according to the procedure of Example 31, Part B.
(C)
rac-5-Fluoro-2-methyl-3-(N-benzyl)-.alpha.-(2-propylamino)-acetamide.
To 5-fluoro-2-methyl-3-(N-2-propyl-N-mesyloxy)-indenylacetamide (2
mmol) in CH.sub.2 Cl.sub.2 (25 ml) at 0.degree. C. is added
benzylamine (4.4 mmol) in CH.sub.2 Cl.sub.2 (15 ml) over a period
of 30 minutes. The resulting solution is stirred at 0.degree. C.
for 1 hour, and at room temperature overnight. The solvent is
removed, and the residue is treated with 1 N NaOH, and is extracted
with CH.sub.2 Cl.sub.2 (100 ml). The extract is washed with water
(2.times.10 ml), and is dried over MgSO.sub.4. After rotary
evaporation, the product is purified by flash chromatography.
(D)
rac-(Z)-5-Fluoro-2-methyl-(4-pyridinylidene)-3-(N-benzyl)-indenyl-.alpha.-
(2-propylamino)-acetamide is obtained from
rac-5-fluoro-2-methyl-3-(N-benzyl)-.alpha.-(2-propylamino)-acetamide
using the procedure of Example 1, Part G (R.sub.1 =F, R.sub.2
=CH.sub.3, R.sub.3 =isopropylamino, R.sub.4 =H, R.sub.5 =H, R.sub.6
=H, R.sub.7 =H, n=1, m=1, Y=4-pyridinyl).
EXAMPLE 34
(Z)-6-Methoxy-2-Methyl-(4-Pyrdinylidene)-3-(N-Benzyl)-Indenylacetamide
(A) Ethyl-2-Hydroxy-2-(p-Methoxyphenyl)-1-Methylpropionate
In a 500 ml. 3-necked flask is placed 36.2 g. (0.55 mole) of zinc
dust, a 250 ml. addition funnel is charged with a solution of 80
ml. anhydrous benzene, 20 ml. of anhydrous ether, 80 g. (0.58 mole)
of p-anisaldehyde and 98 g. (0.55 mole) of ethyl-2-bromoproplonate.
About 10 ml. of the solution is added to the zinc dust with
vigorous stirring, and the mixture is warmed gently until an
exothermic reaction commences. The remainder is added dropwise at
such a rate that the reaction mixture continues to reflux smoothly
(ca. 30-35 min.). After addition is completed the mixture is placed
in a water bath and refluxed for 30 minutes. After cooling to
0.degree., 250 ml. of 10% sulfuric acid is added with vigorous
stirring. The benzene layer is extracted twice with 50 ml. portions
of 5% sulfuric acid and washed twice with 50 ml. portions of water.
The combined aqueous acidic layers are extracted with 2.times.50
ml. ether. The combined etheral and benzene extracts are dried over
sodium sulfate. Evaporation of solvent and fractionation of the
residue through a 6"Vigreux column affords 89 g. (60%) of the
product, ethyl-2-hydroxy-2-(p-methoxyphenyl)-1-methylpropionate,
B.P. 165-160.degree. (1.5 mm.).
(B) 6-Methoxy-2-methylindanone
By the method described in Vander Zanden, Rec. Trav. Chim., 68, 413
(1949), the compound from part A is converted to
6-methoxy-2-methylindanone.
Alternatively, the same compound can be obtained by adding
.alpha.-methyl-p-(p-methoxylphenyl)propionic acid (15 g.) to 170 g.
of polyphosphoric acid at 50.degree. and heating the mixture at
83-90.degree. for two hours. The syrup is poured into iced water.
The mixture is stirred for one-half hour, and is extracted with
ether (3.times.). The etheral solution is washed with water
(2.times.) and 5% NaHCO.sub.3 (5.times.) until all acidic material
has been removed, and is dried over sodium sulfate. Evaporation of
the solution gives 9.1 g. of the indanone as a pale yellow oil.
(C)
(Z)-6-Methoxy-2-methyl-(4-pyridinylidene)-3-(N-benzyl)-indenylacetamide
In accordance with the procedures described in Example 1, parts
D-G, this compound is obtained substituting
6-methoxy-2-methylindanone for 6-fluoro-2-methylindanone in part D
of Example 1.
EXAMPLE 35
(Z)-5-Methoxy-2-Methyl-(4-Pyridinylidene)-3-(N-Benzyl)-Indenylacetamide
(A) Ethyl 5-Methoxy-2-Methyl-3-Indenyl Acetate
A solution of 13.4 g of 6-methoxy-2-methylindanone and 21 g. of
ethyl bromoacetate in 45 ml. benzene is added over a period of five
minutes to 21 g. of zinc amalgam (prepared according to Org. Syn.
Coll. Vol. 3) in 110 ml. benzene and 40 ml. dry ether. A few
cyrstals of iodine are added to start the reaction, and the
reaction mixture is maintained at reflux temperature (ca. 650) with
external heating. At three-hour intervals, two batches of 10 g.
zinc amalgam and 10 g. bromoester are added and the mixture is then
refluxed for 8 hours. After addition of 30 ml. of ethanol and 150
ml. of acetic acid, the mixture is poured into 700 ml. of 50%
aqueous acetic acid. The organic layer is separated, and the
aqueous layer is extracted twice with ether. The combined organic
layers are washed thoroughly with water, ammonium hydroxide and
water. Drying over sodium sulfate, evaporation of solvent in vacuo
followed by pumping at 80.degree. (bath temperature)(l-2 mm.) gives
crude ethyl-(1-hydroxy-2-methyl-6-methoxy-indenyl) acetate (ca. 18
g.).
A mixture of the above crude hydroxyester, 20 g. of
p-toluenesulfonic acid monohydrate and 20 g. of anhydrous calcium
chloride in 250 ml. toluene is refluxed overnight. The solution is
filtered, and the solid residue is washed with toluene. The
combined toluene solution is washed with water, sodium bicarbonate,
water and then dried over sodium sulfate. After evaporation, the
crude ethyl 5-methoxy-2-methyl-3-indenyl acetate is chromatographed
on acid-washed alumina and the product is eluted with petroleum
ether-ether (v./v. 50-100%) as a yellow oil (11.8 g., 70%).
(B)
(Z)-5-Methoxy-2-methyl-(4-pyridinylidene)-3-(N-benzyl)-indenylacetamide
In accordance with the procedures described in Example 1, parts
E-G, this compound is obtained substituting
ethyl-5-methoxy-2-methyl-3-indenyl acetate for
5-fluoro-2-methindenyl-3-acetic acid in Example 1, part E.
EXAMPLE 36
(Z)-.alpha.-5-Methoxy-2-Methyl-(4-Pyridinylidene)-3-(N-Benzyl)-Indenylpropi
onamide
(A) .alpha.-(5-Methoxy-2-methyl-3-indenyl)propionic acid
The procedure of Example 35, part (A) is followed using ethyl
.alpha.-bromopropionate in equivalent quantities in place of ethyl
bromoacetate used therein. There is obtained ethyl
.alpha.-(1-hydroxy-6-methoxy-2-methyl-1-indanyl)propionate, which
is dehydrated to ethyl
.alpha.-(5-methoxy-2-methyl-3-indenyl)propionate in the same
manner.
The above ester is saponified to give
.alpha.-(5-methoxy-2-methyl-3-indenyl)propionic acid.
(B)
(Z)-.alpha.-5-Methoxy-2-methyl-(4-pyridinyl)-3-(N-benzyl)-.alpha.-methyl
i ndenylpropionamide
In accordance with the procedures described in Example 1, parts
E-G, this compound is obtained substituting
.alpha.-5-methoxy-2-methyl-3-indenyl)propionic acid for
5-fluoro-2-methylindenyl-3-acetic acid in Example 1, part E.
EXAMPLE 37
(Z)
.alpha.-Fluoro-5-Methoxy-2-Methyl-(4-Pyridinylidene)-3-(-Benzyl)Indenylace
tamide
(A) Methyl-5-Methoxy-2-Methyl-3-Indenyl-.alpha.-Fluoro Acetate
A mixture of potassium fluoride (0.1 mole) and
methyl-5-methoxy-2-methyl-3-indenyl-.alpha.-tosyloxy acetate (0.05
mole) in 200 ml. dimethylformamide is heated under nitrogen at the
reflux temperature for 2-4 hours. The reaction mixture is cooled,
poured into iced water and then extracted with ether. The ethereal
solution is washed with water, sodium bicarbonate and dried over
sodium sulfate. Evaporation of the solvent and chromatography of
the residue on an acid-washed alumina column (300 g.) using
ether-petroleum ether (v./v. 20-50%) as eluent give the product,
methyl-5-methoxy-2-methyl-3-indenyl-.alpha.-fluoroacetate.
(B) (Z)
.alpha.-Fluoro-5-methoxy-2-methyl-(4-pyridinylidene)-3-(N-benzl)indenylace
tamide
In accordance with the procedures described in Example 1, parts
E-G, this compound is obtained substituting
methyl-5-methoxy-2-methyl-3-indenyl-.alpha.-fluoroacetate for
5-fluoro-2-methylindenyl-3-acetic acid in Example 1, part E.
For the introduction of the .dbd.CH--Y part in Scheme III, any of
the appropriate heterocyclic aldehydes may be used either directly
in the base-catalyzed condensation or in a Wittig reaction in an
alternative route. The aldehydes that may be used are listed in
Table 1 below:
TABLE 1 ______________________________________ pyrrol-2-aldehyde*
pyrimidine-2-aldehyde 6-methylpyridine-2-aldehyde*
1-methylbenzimidazole-2-aldehyde isoquinoline-4-aldehyde
4-pyridinecarboxaldehyde* 3-pyridinecarboxaldehyde*
2-pyridinecarboxaldehyde* 4,6-dimethyl-2-pyridinecarboxaldehyde*
4-methyl-pyridinecarboxaldehyde* 4-quinolinecarboxaldehyde*
3-quinolinecarboxaldehyde* 2-quinolinecarboxaldehyde*
2-chloro-3-quinolinecarboxaldehyde* pyrazinealdehyde (Prepared as
described by Rutner et al., JOC 1963, 28, 1898-99)
pyridazine-3-aldehyde (Prepared as described by Heinisch et al.,
Monatshefte Fuer Chemie 108, 213-224, 1977) pyrimidine-4-aldehyde
(Prepared as described by Bredereck et al., Chem. Ber. 1964, 97,
3407-17) 2-methyl-pyrimidine-4-aldehyde (Prepared as described by
Bredereck et al., Chem. Ber. 1964, 97, 3407-17)
pyridazine-4-aldehyde (Prepared as described by Heinisch et al.,
Monatshefte Fuer Chemie 104, 1372-1382 (1973))
1-methylindole-3-carboxaldehyde* 1-acetyl-3-indolecarboxaldehyde*
______________________________________
The aldehydes above can be used in the reaction schemes above in
combination with various appropriate amines to produce compounds
with the scope of this invention. Examples of appropriate amines
are those listed in Table 2 below:
TABLE 2 ______________________________________ benzylamine
2,4-dimethoxybenzylamine 2-methoxybenzylamine 2-fluorobenzylamine
4-dimethylaminobenzylamine 4-sulfonaminobenzylamine
1-phenylethylamine (R-enantiomer) 2-amino-2-phenylethanol
(S-enantiomer) 2-phenylglycinonitrile (S-enantiomer)
______________________________________
EXAMPLE 38
(Z)-5-Fluoro-2-Methyl-(4-Pyridylidene)-3-(N-Benzyl)
Indenylacetamide Hydrochloride
(Z)-5-Fluoro-2-methyl-(4-pyridylidene)-3-(N-benzyl)indenylacetamide
(1396 g; MW 384.45; 3.63 mol) from Example 1 is dissolved at
45.degree. C. in ethanol (28 L). Aqueous HCl (12 M; 363 mL) is
added stepwise. The reaction mixture is heated under reflux for 1
hour, is allowed to cool to room temperature, then stored at
-10.degree. C. for 3 hours. The resulting solid is filtered off, is
washed with ether (2.times.1.5 L) and is air-dried overnight.
Drying under vacuum at 70.degree. C. for 3 days gives
(Z)-5-fluoro-2-methyl-(4-pyridylidene)-3-(N-benzyl)indenylacetamide
hydrochloride with a melting point of 207-209.degree. C. (R.sub.1
=F, R.sub.2 =CH.sub.3, R.sub.3 =H, R.sub.4 =H, R.sub.5 =H, R.sub.6
=H, R.sub.7 =H, n=1, m=1, Y=4-pyridinyl.multidot.hydrochloride).
Yield: 1481 g (97%; 3.51 mol); MW: 420.91 g/mol.
.sup.1 H-NMR (DMSO-d.sub.6): 2.18 (s,3,.dbd.C--CH.sub.3); 3.54
(s,2,.dbd.CH.sub.2 CO); 4.28 (d,2,NCH.sub.2); 6.71 (m,1,ar.); 7.17
(m,8,ar.); 8.11 (d,2,ar., AB system); 8.85 (m,1,NH); 8.95
(d,2,ar.,AB system); IR (KBr): 3432 NH; 1635 C.dbd.O; 1598
C.dbd.C.
Biological Effects
(A) Growth Inhibition
The compound of Example 1 was assayed for its growth inhibitory
activity on the human colon carcinoma cell line, SW-480 obtained
from ATCC (Rockville, Md.), to ascertain the degree of growth
inhibition. Growth inhibition of this cell line is indicative of a
benefit on precancerous lesions and neoplasms. The cell line and
growth assay employed for such experiments are well characterized,
and are used to evaluate the anti-neoplastic properties of NSAIDs.
The assay is used by the United States National Cancer Institute in
its screening program for new anti-cancer drugs.
Drug stock solutions were made in 100% DMSO and were then diluted
with RPMI media for cell culture testing. All drug solutions were
prepared fresh on the day of testing. The cultured cells were
obtained at passage #99 and grown in RPMI media supplemented with
5% fetal calf serum, and 2 mM glutamine, 100 U/ml penicillin, 100
U/ml streptomycin, and 0.25 .mu.g/ml amphotericin. The cultures
were maintained in a humidified atmosphere of 95% air and 5%
CO.sub.2 at 37.degree. C. The cultures were passaged at
preconfluent densities using a solution of 0.05% trypsin and 0.53
mM EDTA. Cells were plated at 1000 cells/well for 96 well
flat-bottom microtiter plates.
Tumor cell growth inhibition was assessed using the Sulforhodamine
B (SRB) protein binding assay. In this assay, tumor cells were
plated in 96-well plates and treated with drug-containing media for
six days (continuous exposure). For each plate, 6 wells were
designated as no treatment controls, six wells as vehicle (0.1%
DMSO) controls, and the remaining wells for drug dilutions with
three wells per drug concentration. At the end of the exposure
period, the cells were fixed and stained with sulforhodamine B, a
protein binding dye. The dye was then solubilized, and the optical
density of the resulting solution was determined on a 96-well plate
reader. The mean dye intensity of the treated wells was then
divided by the mean dye intensity in the control wells (6 wells of
each) to determine the effect of the drug on the cells. Dye
intensity is proportional to the number of cells or amount of
protein per well. The resultant "percent inhibition" value then
represents the degree of growth inhibition caused by the drug.
For each experiment, an IC.sub.50 value was determined and used for
comparative purposes. This value is equivalent to the concentration
of drug needed to inhibit tumor cell growth by 50%. IC.sub.50 value
was obtained graphically by connecting the mean values for each
drug concentration tested. Each experiment included at least three
wells per drug concentration. Concentration was plotted on a log
scale on the X-axis. IC.sub.50 value obtained for the compound of
Example 1wa 0.724 for the SW-480 cell line.
(B) Cyclooxygenase (COX) Inhibition
COX catalyzes the formation of prostaglandins and thromboxane by
the oxidative metabolism of arachidonic acid. The compound of
Example 1 of this invention, as well as a positive control,
(sulindac sulfide) were evaluated to determine whether they
inhibited purified cyclooxygenase Type I (see Table 3 below).
The compounds of this invention were evaluated for inhibitory
effects on purified COX. The COX was purified from ram seminal
vesicles, as described by Boopathy, R. and Balasubramanian, J.,
239:371-377, 1988. COX activity was assayed as described by Evans,
A. T., et al., "Actions of Cannabis Constituents on Enzymes Of
Arachidonate Metabolism Anti-Inflammatory Potential," Biochem.
Pharmacol., 36:2035-2037, 1987. Briefly, purified COX was incubated
with arachidonic acid (100 .mu.M) for 2.0 min at 37.degree. C. in
the presence or absence of test compounds. The assay was terminated
by the addition of TCA, and COX activity was determined by
absorbance at 530 nm.
TABLE 3 ______________________________________ COX I EXAMPLE %
Inhibition (100 .mu.M) ______________________________________ (*
-1000-M) Sulindac sulfide 86 1 <25
______________________________________
(C) Apoptsis
Apoptosis was measured using an assay of cell death based on
morphological characteristics of apoptotic cells (i. e., condensed
chromatin). Drug preparation and cell culture conditions were the
same as for the SRB assay described above, except that HT-29 human
colon carcinoma cells were used. Confluent cultures were
established in 12.5 cm.sup.2 flasks by plating 0.5.times.10.sup.6
cells/flask. The cultures were assayed for apoptosis by fluorescent
microscopy following labeling with acridine orange and ethidium
bromide. Floating and attached cells were collected by
trypsinization and washed three times in PBS. One ml aliquots were
centrifuged (3 g). The pellet was resuspended in 25 .mu.l media and
1 .mu.l of a dye mixture containing 100 .mu.g/ml acridine orange
and 100 .mu.g/ml ethidium bromide prepared in PBS and mixed gently.
Ten .mu.l of the mixture was placed on a microscope slide and
covered with a 22 mm.sup.2 coverslip, was examined with 40x dry
objectives under epillumination by filter combination.
An observer blinded in regard to the identity of the samples scored
at least 100 cells per sample. Apoptotic cells were identified by
nuclear condensation of chromatin stained by the acridine orange or
ethidium bromide. These results are provided in Table 4 below.
TABLE 4 ______________________________________ Apoptosis Effects of
Compounds Morphology % Apoptotic Cells DNA Fragmentation EXAMPLE (1
.mu.M) FS (100 .mu.M) EC.sub.50 (.mu.M)
______________________________________ 1 88 4.2 29 2 5.4 3 8.5 4
3.9 38 15 ______________________________________
Apoptosis was also measured based on the amount of fragmented DNA
contained in cell lysates. Briefly, SW-480 colon adenocarcinoma
cells were plated in 96-well microtitre plates ("MTP") at a density
of 10K cells/well in 180.mu.1 and were incubated for 24 hrs. Cells
were then treated with 20.mu.1 aliquots of appropriately diluted
compound, and allowed to incubate for an additional 48 hrs.
After the incubation, samples were prepared according to the
following steps. The MTP was centrifuged (15 min., 1000 rpm) and
the supernatant was carefully removed by fast inversion of the MTP.
The cell pellets in each well were resuspended in 200 .mu.l lysis
buffer and incubated for 45 min. at room temperature to lyse the
cells. The lysates were then centrifuged (15 min., 1000 rpm) and 20
.mu.l aliquots of the supernatant (=cytoplasmic fraction) were
transferred into the streptavidin coated MTP for analysis. Care was
taken not to shake the lysed pellets in the MTP (=cell nucleii
containing high molecular weight, unfragmented DNA). Samples were
analyzed immediately, because storage at 4 C or -20.degree. C.
reduces the ELISA-signals.
Samples were then processed according to a DNA fragmentation assay
protocol, and dose-response curves were generated based on optical
density readings. Quantification of DNA was done by a commercially
available photometric enzyme-immunoassay manufactured by
Mannheim-Boehringer under the name "Cell Death Detection
ELISA.sup.plus ". The assay is based on a quantitative
sandwich-enzyme-immunoassay-principle using mouse monoclonal
antibodies directed against DNA and histones, respectively. This
allows the specific determination of mono and oligonucleosomes in
the cytoplasmatic fraction of cell lysates. In brief, the assay
procedure is as follows. The sample (cell-lysate, serum,
culture-supernatant etc.) is placed into a streptavidin-coated MTP.
Subsequently, a mixture of anti-histone-biotin and anti-DNA-POD is
followed by incubation for 2 hours. During the incubation period,
the anti-histone antibody binds to the histone-component of the
nucleosomes and simultaneously fixes the immunocomplex to the
streptavidin-coated MTP via its biotinylation. Additionally, the
anti-DNA-POD antibody reacts with the DNA component of the
nucleosomes. After removal of unbound antibodies by a washing step,
the amount of nucleosomes is quantified by the POD retained in the
immunocomplex. POD is determined photometrically with ABTS.RTM.
(2,2'-Azino-di[3-ethylbenzthiazolin-sulfonat])* as substrate.
Fold stimulation (FS=ODmax/ODveh), an indicator of apoptotic
response, was determined for each compound tested. EC.sub.50 values
were determined either specifically by data analysis software, or
by estimates based on the effective concentration range of each
compound (ECR=min. effective dose-min. dose to peak effect). These
FS and EC.sub.50 values for the tested compounds are listed above
in Table 4.
In addition, using the DNA fragmentation test above, a dose
response for the compound of Example 1 was obtained. Those data are
set forth in Table 5.
TABLE 5 ______________________________________ Apoptosis Level Dose
(.mu.M) (Mean OD Value .+-. SD)
______________________________________ 0.5 0.186 .+-. 0.008 1.0
0.207 .+-. 0.061 5.0 0.208 .+-. 0.073 10 0.296 .+-. 0.050 50 0.500
.+-. 0.048 100 0.633 .+-. 0.053 500 0.659 .+-. 0.012
______________________________________
The compounds of this invention can be formulated with
pharmaceutically acceptable carriers into unit dosage forms in a
conventional manner so that the patient in need of therapy for
precancerous lesions can periodically (e.g., once or more per day)
take a compound according to the methods of this invention. The
exact initial dose of the compounds of this invention can be
determined with reasonable experimentation. One skilled in the art
should understand that the initial dosage should be sufficient to
achieve a blood plasma concentration approaching a percentage of
the IC.sub.50 value of the compound, with the percentage depending
on the chemopreventative or chemotherapeutic indication. The
initial dosage calculation would also take into consideration
several factors, such as the formulation and mode of
administration, e.g. oral or intravenous, of the particular
compound. For example, assuming a patient with an average
circulatory system volume of about four liters, based on the
IC.sub.50 values for compounds of this invention, one would
calculate a dosage of about 0.6 mg-4.0 gr of such compounds for
intravenous administration to achieve a systemic circulatory
concentration equivalent to the IC.sub.50 concentration.
Compounds of this invention are also cGMP-specific PDE inhibitors
as taught in U.S. patent application Ser. No. 09/046,739 filed Mar.
24, 1998. Compounds can be tested for inhibitory effect on
phosphodiesterase activity using either the enzyme isolated from
any tumor cell line such as HT-29 or SW-480. Phosphodiesterase
activity can be determined using methods known in the art, such as
a method using radioactive .sup.3 H cyclic GMP (cGMP)(cyclic
3',5'-guanosine monophosphate) as the substrate for PDE5 enzyme.
(Thompson, W. J., Teraski, W. L., Epstein, P. M., Strada, S. J.,
Advances in Cyclic Nucleotide Research, 10:69-92, 1979, which is
incorporated herein by reference). In brief, a solution of defined
substrate .sup.3 H-cGMP specific activity (0.2 .mu.M; 100,000 cpm;
containing 40 mM Tris-HCl (pH 8.0), 5 mM MgCl.sub.2 and 1 mg/ml
BSA) is mixed with the drug to be tested in a total volume of 400
.mu.l. The mixture is incubated at 30.degree. C. for 10 minutes
with partially purified cGMP-specific PDE isolated from HT-29
cells. Reactions are terminated, for example, by boiling the
reaction mixture for 75 seconds. After cooling on ice, 100 .mu.l of
0.5 mg/ml snake venom (O. Hannah venom available from Sigma) is
added and incubated for 10 min at 30.degree. C. This reaction is
then terminated by the addition of an alcohol, e.g. 1 ml of 100%
methanol. Assay samples are applied to a anion chromatography
column (1 ml Dowex, from Aldrich) and washed with 1 ml of 100%
methanol. The amount of radioactivity in the breakthrough and the
wash from the columns in then measured with a scintillation
counter. The degree of PDE5 inhibition is determined by calculating
the amount of radioactivity in drug-treated reactions and comparing
against a control sample (a reaction mixture lacking the tested
compound).
Using such protocols, the cGMP-specific PDE inhibitor of Example 1
had an IC.sub.50 value of 0.68 .mu.M utilizing HT29 cell
extracts.
It will be understood that various changes and modifications can be
made in the details of procedure, formulation and use without
departing from the spirit of the invention, especially as defined
in the following claims.
* * * * *